Kovacs Zsuzsanna, Simon Adam, Szabo Zoltan, Nagy Zsolt, Varoczy Laszlo, Pal Ildiko, Csanky Eszter, Guttman Andras
Horváth Csaba Memorial Institute for Bioanalytical Research, University of Debrecen, Hungary.
Thermo Fisher Scientific, Sunnyvale, CA, USA.
Electrophoresis. 2017 Sep;38(17):2115-2123. doi: 10.1002/elps.201700006. Epub 2017 Feb 21.
Multiple myeloma (MM) is an immedicable malignancy of the human plasma cells producing abnormal antibodies (also referred to as paraproteins) leading to kidney problems and hyperviscosity syndrome. In this paper, we report on the N-glycosylation analysis of paraproteins from total human serum as well as the fragment crystallizable region (F ) and fragment antigen binding (F ) κ/λ light chain fractions of papain digested immunoglobulins from multiple myeloma patients. CE-LIF detection was used for the analysis of the N-glycans after endoglycosidase (PNGase F) mediated sugar release and fluorophore labeling (APTS). While characteristic N-glycosylation pattern differences were found between normal control and untreated, treated and remission stage multiple myeloma patient samples at the global serum level, less distinctive changes were observed at the immunoglobulin level. Principal component analysis adequately differentiated the four groups (control and three patient groups) on the basis of total serum N-glycosylation analysis. 12 N-glycan features showed statistically significant differences (p <0.05) among various stages of the disease in comparison to the control at the serum level, while only six features were identified with similar significance at the immunoglobulin level, including the analysis of the partitioned F fragment as well as the F κ and F λ chains.
多发性骨髓瘤（MM）是一种无法治愈的人类浆细胞恶性肿瘤，会产生异常抗体（也称为副蛋白），导致肾脏问题和高黏滞综合征。在本文中，我们报告了对来自人全血清的副蛋白以及来自多发性骨髓瘤患者经木瓜蛋白酶消化的免疫球蛋白的可结晶片段（Fc）和抗原结合片段（Fab）κ/λ轻链组分的N-糖基化分析。在糖苷内切酶（PNGase F）介导的糖释放和荧光团标记（APTS）后，采用毛细管电泳-激光诱导荧光检测（CE-LIF）对N-聚糖进行分析。虽然在全球血清水平上，正常对照与未治疗、治疗和缓解期多发性骨髓瘤患者样本之间发现了特征性的N-糖基化模式差异，但在免疫球蛋白水平上观察到的差异不太明显。主成分分析基于全血清N-糖基化分析充分区分了四组（对照组和三个患者组）。与对照组相比，在血清水平上，12种N-聚糖特征在疾病的各个阶段显示出统计学上的显著差异（p<0.05），而在免疫球蛋白水平上只有6种特征具有类似的显著性，包括对分离的Fc片段以及Fabκ和Fabλ链的分析。
